Bioreactor paradigm for the production of secondary extra-particle hyphal matrices

ABSTRACT

The invention describes a methodology for production of a secondary extra-particle fungal matrix for application as a mycological material, manufactured via a Type II actively aerated static packed-bed bioreactor. A pre-conditioned air stream is passed through a substrate of discrete elements inoculated with a filamentous fungus to form an isotropic inter-particle hyphal matrix between the discrete elements. Continued feeding of the air through the substrate of discrete elements and isotropic inter-particle hyphal matrixes develops an extra-particle hyphal matrix that extends from an isotropic inter-particle hyphal matrix in the direction of airflow into a void space within the vessel.

INCORPORATION BY REFERENCE TO ANY PRIORITY APPLICATIONS

This application is a continuation application of U.S. patentapplication Ser. No. 16/549,757, filed Aug. 23, 2019, which claims thebenefit of U.S. Provisional Patent Application 62/740,159, filed Oct. 2,2018, the entireties of which are hereby expressly incorporated byreference herein.

BACKGROUND OF THE INVENTION Field of the Invention

This invention relates to a bioreactor paradigm for the production ofsecondary extra-particle hyphal matrices. More particularly, thisinvention relates to an actively aerated packed-bed bioreactor paradigmfor the production of secondary extra-particle hyphal matrices. Stillmore particularly, this invention relates to a method of producing amycological material.

Description of the Related Art

As is known from published US Patent Application 2015/0033620, amycological biopolymer product consisting entirely of fungal myceliummay be made by inoculating a nutritive substrate with a selected fungusin a sealed environment except for a void space, which space issubsequently filled with a network of fungal mycelium. The environmentalconditions for producing the mycological biopolymer product, i.e., ahigh carbon dioxide (CO2) content i.e., from 5% to 7% by volume and anelevated temperature i.e., from 85° F. to 95° F., prevent fulldifferentiation of the fungus into a mushroom. There are no stipe, cap,or spores produced. The biopolymer product grows into the void space ofthe tool, filling the space with an undifferentiated myceliumchitin-polymer, which is subsequently extracted from the substrate anddried.

As is also known from pending U.S. patent application Ser. No.16/190,585, filed Nov. 14, 2018, another method of growing a biopolymermaterial employs incubation of a growth media comprised of nutritivesubstrate and a fungus in containers that are placed in a closedincubation chamber with air flows passed over each container while thechamber is maintained with a predetermined environment of humidity,temperature, carbon dioxide and oxygen. The mycological biopolymer isgrown into a panel at a dry density of 0.5 to 4 pounds per cubic foot ona dry mass basis.

Filamentous fungi are comprised of cross-linked networks of filamentouscells called hyphae, which expand via polarized tip extension and branchformation (increasing the number of growing tips), which is equivalentto cell division in animals and plants. Hyphal tip extension can displaya number of tropisms (positive or negative) including gravitropisms,autotropisms, and galvanotropisms, of which modification is adequate toaffect meaningful organizational and morphological variety in the fungalthallus (mycelium) and fruiting bodies (mushrooms).

Filamentous fungi are defined by their phenotypic plasticity and mayproduce a secondary mycelium which, based on the “fuzzy logic” ofdifferentiation as a function of differential expression of discrete“subroutines” rather than defined pathways, can express variable degreesof differentiation spanning from complex reproductive structures(mushrooms) to a completely undifferentiated vegetative mycelium.

Type I unaerated and unmixed bioreactors represent one of the mostcommonly used paradigms for solid-state fermentation, which consists ofa shallow tray containing solid-substrate and fungal precursor, which isplaced in either an active or passively controlled incubationenvironment where fungal growth is dependent on passive oxygen and heatdiffusion between the fungal-particle matrix and the externalenvironment. Within this paradigm, the depth of the tray represents theprimary limiting variable affecting oxygen and temperature diffusion.

As described in published US Patent Application 2018/0146627, theexpansion and isolation of a secondary extra-particle mycelium from asolid-substrate fermented with filamentous fungi is specificallydependent on Type I tray-based bioreactor systems. In these systems,passive diffusion of respiratory effluent is used to determine thegrowth environment, within which expansion of the secondaryextra-particle mycelium occurs as a function of negatively gravitropicexpansion. Within this paradigm, even if the tray is maintained withinan actively controlled incubation environment, temperature and gasexchange inputs remain inherently indirect as oxygen and temperaturediffusion will remain passive at the interface of the fungal-particlematrix and external environments. Additionally, the environment withinthe developing extra-particle mycelium is controlled only throughdiffusion, which becomes an increasing restriction as the thickness anddensity of the extra-particle mycelium increases. This feedback imposeslogical limitations on large scale development of tissue and mayincrease morphological heterogeneity.

Type II actively aerated and unmixed bioreactors represent a class ofbioreactor defined by a static bed of discrete particles, around andbetween which microbial fermentation occurs, with air activelyintroduced from either end of the particle bed with forced diffusionthrough the particle matrix. This class of bioreactor allows for activeheat removal and supply of oxygen through the particle bed, reducing oreliminating bed depth as a significant limiting variable. Further, theair stream can be pre-conditioned to specific temperatures prior toinput into the particle bed, allowing for modification of the directtemperature and gas exchange rates experienced by the inter-particleenvironment during fermentation, and further allowing for specifictemporal modification of these variables.

Accordingly, it is an object of the invention to efficiently use a TypeII actively aerated static packed-bed bioreactor to manufacture asecondary extra-particle fungal matrix for application as a mycologicalmaterial.

It is another object of the invention to provide for a greater degree ofmaterial morphology and property control in the manufacture of asecondary extra-particle fungal matrix.

It is another object of the invention to simplify the manufacture of amycological material.

Briefly, the invention provides a method of producing a mycologicalmaterial employing a Type II actively aerated static packed-bedbioreactor.

The method includes the steps of providing a vessel having a chamber andloading a substrate of discrete elements inoculated with a filamentousfungus into the chamber.

The vessel is characterized in being constructed to contain the loadedsubstrate in a condition wherein an air stream can be passed through thesubstrate and out of the vessel. In one embodiment, the vessel may beoriented vertically and the air stream passed vertically through thesubstrate either upwardly or downwardly. In another embodiment, thevessel may be oriented horizontally, and the air stream passedhorizontally through the substrate.

In still another embodiment, the vessel is characterized in beingconstructed to contain the loaded substrate in vertically orhorizontally separated sections with the air stream introduced betweenthe separated sections of substrate to flow through each of thesections.

In operation, after loading of the vessel, a pre-conditioned air streamis fed through the vessel for diffusion between the discrete elements inthe chamber and for a time sufficient for the filamentous fungus toexpand a contiguous network of hyphae between and around the discreteelements to form an isotropic inter-particle hyphal matrix (IPM).

In accordance with the method, the pre-conditioned air stream continuesto be fed through the vessel for diffusion between the discrete elementsand the isotropic inter-particle hyphal matrix for a time sufficient todevelop a polarized condition within the vessel wherein air exits theisotropic inter-particle hyphal matrix as a laminar flow into at leastone void space within the vessel and to form an extra-particle hyphalmatrix (EPM) extending from the isotropic inter-particle hyphal matrixin the direction of airflow within the at least one void space.

The discrete elements may be in the form of particles that can supportfilamentous fungal growth. For example, the particles may belignocellulose (e.g., agricultural residue, wood), which would act as anutrient source for the fungus. The particles could be acquired andprocessed according to typical known processes; for instance, harvestedfrom trees and ground to size using a hammer mill

Alternatively, the particles may be ones that do not act as a nutrientsource for the fungus but only as a solid support for supplementednutrition and fungal growth (for example, pearlite mixed with water andsupplemented nutrients to support fungal growth).

The discrete elements may also be in the form of fibers so long as aircan be passed through the fiber matrix and filamentous fungal growth canoccur around and between the fibers.

In one embodiment, the vessel has a permeable partition within thevessel to separate the chamber with the loaded inoculated substrate froma void space and the air stream is fed through the vessel to passdownwardly through the chamber into the void space. The vessel should bepermeable on the exit end in order to allow the effluent air stream toexit the void space of the vessel. This could be done by perforating thevessel end, through a valve, or any other means of allowing effluent airto escape.

In a second embodiment, the air stream is fed through the vessel to passupwardly through the chamber with the loaded inoculated substrate into avoid space above the loaded chamber.

In a third embodiment, the substrate of discrete elements inoculatedwith a filamentous fungus is separated into two spaced apart sectionswithin the chamber of the vessel and the pre-conditioned air stream isfed into the vessel between the substrate sections for diffusion betweenthe discrete elements in each section to form an isotropicinter-particle hyphal matrix therein and to form an extra-particlehyphal matrix extending from the isotropic inter-particle hyphal matrixin the direction of airflow.

This latter embodiment produces two separate extra-particle hyphalmatrixes; one at one end of the vessel and another at the opposite endof the vessel.

In each embodiment, the vessel may be disposed vertically so that theair stream flows vertically through the inoculated substrate andisotropic inter-particle hyphal matrix, or the vessel may be disposedhorizontally so that the air stream flows horizontally through theinoculated substrate and isotropic inter-particle hyphal matrix.

In a fifth embodiment, which is applicable to any of the above fourembodiments, at least one of a paramorphogen (such as terpene or alkylpyrone), a volatile compound, and other aromatic compounds may beintroduced into the air prior to permeation through the substrateparticle matrix, IPM, and EPM in order to further modify the specificmorphology and density of the EPM.

In a sixth embodiment, which is applicable to any of the above fiveembodiments, a second low-density substrate, which need not be suitablefor supporting IPM on its own, is included in the void space. Duringmanufacture, EPM extends from the inoculated matrix as it would into anopen space, but instead grows around and within the low-density secondsubstrate generating a composite of EPM and secondary substrate withmodified material properties.

In a seventh embodiment, which is applicable to any of the above sixembodiments, the bottom void space is of a defined geometry. Duringmanufacture, EPM 8 expands into the bottom void space creating an EPM ofthe same geometry as the bottom void space.

Critically, Type I tray bioreactor systems depend on either ahomogenized or heterogeneous extra-matrix environment (i.e., external ofthe particle-fungal matrix) developed as a function of passiverespiratory effluent dissipation, with gas exchange and heat diffusionoccurring passively at the interface of the extra- and inter-particlematrix environments. Published US Patent Application 2015/0033620teaches specifically to regulation of CO2 to no less than 3%, withregular exhausting to manage this gas concentration.

Further, the prior art also teaches explicitly to development of anegatively gravitropic secondary extra-particle mycelium, in whichexpansion only occurs against gravity and into the extra-matrixenvironment. The extra-matrix environment described in the prior art mayrequire active control of the gas concentration.

SUMMARY OF THE INVENTION

The described invention herein leverages a Type II actively aeratedbioreactor paradigm in which air and temperature inputs are inherentlydirect, in that passage of these inputs occurs directly through thesubstrate particle matrix, IPM, and EPM at a defined volumetric (eitherconstant or dynamic) rate. In this case, under most conditions, theoutput respiratory effluent would be <3% CO2, directly teaching againstPublished US Patent Application 2015/0033620.

Filamentous fungi are defined by their phenotypic plasticity, and thetemporal development, morphology, cell concentration, efficiency ofproducing an EPM, and the morphology of the EPM is directly dependent ongas and temperature conditions. Considering this phenotypic plasticity,the direct nature of input conditions (volumetric air exchange rate, airtemperature, introduction of paramorphogens/volatile compounds intoinput air) by the described invention therefore provides for a paradigmfor more direct modification of the morphological characteristics of thedeveloped EPM, and thereby the mechanical properties of the EPM, thanprior art.

Additionally, conditioning of airflow and gas concentration to a stagein which development of EPM occurs is a direct function of the IPM(diffusion through, as well as respiratory effluent from the IPM, whichis modified simply by changing the volumetric air exchange rate anddepth of the substrate particle bed), which allows for considerablesimplification of the bioreactor design as compared to prior art as theneed for elaborate airflow systems and active gas composition controlare reduced, thereby enjoying an increase in scalability for commercialmanufacture. Within the polarized and direct conditions described,development of the EPM may occur as either positively or negativelygravitropic, but will always occur at the output of respiratory effluentfrom the substrate particle matrix-IPM as hyphal extension in thedirection of air flow.

BRIEF DESCRIPTION OF THE DRAWINGS

These and other objects and advantages of the invention will become moreapparent from the following detailed description taken in conjunctionwith the drawings wherein:

FIG. 1A schematically illustrates the direction and pattern of airflowand respiratory effluent through an inoculated substrate in a vessel inaccordance with the method of the invention;

FIG. 1B schematically illustrates the pattern of fungal growth withinthe inoculated substrate of FIG. 1A in accordance with the method of theinvention;

FIG. 2 schematically illustrates an embodiment wherein the direction ofairflow is upward through the inoculated substrate;

FIG. 3 schematically illustrates an embodiment where the substrate ofdiscrete elements inoculated with a filamentous fungus is separated intotwo spaced apart sections within the chamber of a vessel and air passedthrough each section in accordance with the method of the invention;

FIG. 4 schematically illustrates a vessel as in FIG. 3 disposed in ahorizontal manner in accordance with the invention;

FIG. 5 schematically illustrates a vessel of cubic shape for performingthe method of the invention; and

FIG. 6 schematically illustrates a vessel as in FIG. 1A with bottom voidspace of a defined geometry in accordance with the invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

Referring to FIG. 1A, the method of producing a mycological materialcomprising the steps of providing a vessel 1 having a chamber that canbe loaded with a substrate of discrete elements 3 inoculated with afilamentous fungus into the chamber.

As illustrated, the vessel 1 has a head space 2 at the upper end and apermeable partition 5 within the vessel 1 separating the chamber from avoid space 6 below the partition 5.

Pre-conditioned air (at near-saturation and a defined temperature andgas composition) is fed into the top of the vessel 1 (or head-space 2)and diffuses down and between the discrete substrate elements 3 asindicated by the arrows 4 with the air flow exiting through thepermeable partition 5. In this case, the specific gas composition andvolumetric air flow rate may be constant or may be modulateddynamically.

Referring to FIG. 1B, under these conditions, the filamentous fungusexpands a contiguous network of filamentous cells (hyphae) between andaround the discrete substrate particles 3 forming an isotropicinter-particle hyphal matrix (IPM) 7.

As air diffuses between the particles 3 and through the IPM 7, apolarized condition develops within the vessel 1 in which air exitingthe IPM 7 as laminar flow (as a function of the substrate particlematrix-IPM acting as a plenum) into the underlying void space 6 is ofhigher concentration of moisture than air entering the vessel (due tore-saturation during passage through IPM) and contains respiratoryeffluent (CO₂, VOC, other signaling chemicals). Importantly, thiscreates a single, vertically oriented gradient of moisture andrespiratory effluent (a polarized condition) culminating in theunderlying void space 6 experiencing the highest concentrations witheven, laminar flow of the air and respiratory effluent. Within thispolarized condition, an extra-particle hyphal matrix (EPM) 8 extendsfrom the IPM 7 in a positively gravitropic orientation, extending in thedirection of airflow within the bottom void-space 6.

The morphology of EPM 8 is of increased anisotropy as compared to IPM 7with dominant directionality occurring in the vertical orientation. TheEPM 8 is then removed from the permeable partition 5 for utilization asa mycological material per Applications.

The following example is given with respect to FIG. 1A and FIG. 1B.

Example 1. Production of EPM

-   -   1. Discrete lignocellulose particles are amended with        supplemental nutrition, hydrated to a stage amenable to fungal        growth, and combined with the spawn of filamentous fungal        species Ganoderma tsugae (i.e., preparation of inoculated        substrate) to form discrete particles 3 inoculated with a        filamentous fungus.    -   2. Inoculated substrate of inoculated particles 3 are loaded        into the bioreactor vessel 1 which contains a permeable        partition 5 and which consists of a top head-space 2, a chamber        to receive the inoculated substrate matrix, and a bottom        void-space 6 below the permeable partition 5.    -   3. Air is fed into the top (head-space) of the bioreactor vessel        at a rate of 0.2 volumes per bioreactor volume per minute, which        has been pre-conditioned to an average temperature of 85° F. and        a relative humidity (RH) of >90%. This pre-conditioned air        diffuses through the inoculated substrate as indicated by the        arrows 4 and exits through the permeable partition 5, into the        bottom void-space 6, and out of the bioreactor with effluent CO₂        concentration of <3%. These input conditions are maintained for        the duration of the growth cycle.    -   4. Fungal growth occurs within the lignocellulose particle        matrix by development of an isotropic hyphal matrix between and        around the discrete lignocellulose particles (i.e., development        of IPM). As growth of IPM progresses, the flow of        pre-conditioned air per step 3 continues through the IPM,        re-saturating the air to approach 100% RH and evacuating        respiratory effluent, creating a top-down gradient of RH and        respiratory effluent, and laminar flow from the bottom of the        lignocellulose particle matrix-IPM into the bottom void-space 6        and out of the bioreactor.    -   5. From the IPM, a positively gravitropic extra-particle hyphal        matrix (EPM) extends through the permeable partition 5 and into        the bottom void-space 6, extending in the direction of airflow.        The developed EPM represents a distinct structural morphology        from the IPM, with a cell volume density (cell volume per total        volume) of 2× that of the IPM, a directional coherency (degree        of anisotropy of the hyphal matrix) of 3.2× that of IPM, and        oriented hyphal agglomeration (galvanotropism) into strands        increasing the average strand thickness to 1.11× that of IPM.    -   6. EPM is expanded to a target thickness based on the specific        application requirements, then separated from the permeable        partition 5 for post-processing dictated by the specific        application.

Referring to FIG. 2 , wherein like reference characters indicate likeparts as above, the vessel 1 is constructed so that the pre-conditionedair is fed into the bottom of the vessel 1 and diffuses up and betweenthe discrete substrate elements 3 to form isotropic inter-particlehyphal matrixes (IPM) 7 between the elements 3. In this embodiment, theextra-particle hyphal matrixes (EPM) 8 extends from the IPM 7 in thedirection of airflow within the upper void-space 6 and development ofthe EPM is negatively gravitropic.

The following example is given with respect to FIG. 2 .

Example 2. The Procedure of Example 1, Modifying EPM StructuralCharacteristics by Specific Modification of Input Temperature andAirflow Conditions

-   -   1. Example 1 steps 1 and 2.    -   2. Per Example 2 step 3, with average temperature modified to        90° F., and airflow rate modified to 1.2 volumes per bioreactor        volume per minute.    -   3. Example 1 step 4.    -   4. Example 1 step 5, with EPM morphology modified to a cell        volume density of 4.5× that of IPM, a directional coherency of        2.6× that of IPM, and average hyphal strand thickness of 1.29×        that of IPM.    -   5. Example 1 step 6.

Referring to FIG. 3 , wherein like reference characters indicate likeparts as above, the vessel 1 has a pair of permeable partitions 5 atmid-height to form a head space 2 therebetween and a pair of chambersfor loading of two separate sections of the substrate of inoculatedelements 3 therein.

In operation, air is input into the head-space 2 in the center of thebioreactor vessel 1 defined by the permeable partitions 5, from whichair diffuses both down and up through the substrate particle matrix-IPM,in which laminar flow of the respiratory effluent outputs at both thebottom and top of the substrate particle matrix-IPM, where EPM 6manifests as both positively and negatively gravitropic growth.

The following example is given with respect to FIG. 3 .

Example 3. The Procedure of Example 1. Modifying EPM StructuralCharacteristics by Introduction of an Aromatic Compound into the InputAir

-   -   1. Example 1 steps 1 and 2.    -   2. Example 1 step 3, with terpene introduced into the        pre-conditioned air prior to introduction to the inoculated        substrate.    -   3. Example 1 step 4.    -   4. Example 1 step 5, wherein the EPM and/or IPM is of reduced        density and greater directional coherency as a function of the        terpene exposure during IPM/EPM development.    -   5. Example 1 step 6.

Referring to FIG. 4 , wherein like reference characters indicate likeparts as above, the orientation of the vessel 1 of FIG. 3 is adjusted sothat airflow, the gradient of respiratory effluent, and EPM extensionoccurs in the horizontal direction rather than the vertical direction.

Referring to FIG. 5 , wherein like reference characters indicate likeparts as above, the vessel 1 is a 4×4×4fl Type II packed-bed activelyaerated bioreactor of cubic shape.

The following example is given with respect to FIG. 5 .

Example 4. Production of EPM Using a 4×4×4fl Type II Packed-Bed ActivelyAerated Bioreactor

-   -   1. The vessel 1 is a 4×4×4fl container.    -   2. The permeable partition 5 is placed at a depth of 3 ft,        allowing for 1 ft of empty volume 6 below the permeable        partition 5.    -   3. Inoculated substrate elements 3 are loaded in the top 3 ft of        the vessel 1.    -   4. Air is fed into the top (head-space) of the bioreactor vessel        and through the particle matrix per Example 1 steps 3. and 4.    -   5. IPM 7 develops around and between substrate elements 3.    -   6. EPM 8 extends in a positively gravitropic orientation into        the bottom void space 6 to a given target thickness, e.g., a        thickness of up to 12 inches as a function of incubation time.    -   7. EPM is separated from the permeable partition 5 and        post-processed per Example 1 step 6.

Referring to FIG. 6 , wherein like reference characters indicate likeparts as above, the vessel 1 may be made with a base that defines a voidspace 6 of a selected geometric shape, for example, of an ovalcross-sectional shape.

In operation, EPM 8 expands in a positively gravitropic orientation intothe bottom void space 6, producing an EPM of a defined geometry.

Alternatively, a vessel 1 with a base that defines a void space 6 of aselected geometric shape may have a second low-density substratepositioned in the void space 6 and, during operation, the extra-particlehyphal matrix is allowed to grow around and within the secondlow-density substrate to form a composite of the extra-particle hyphalmatrix and the second low-density substrate.

Example 5. Expansion of EPM into a Secondary Substrate to Form anEPM-Secondary Substrate Composite

-   -   1. Example 1 step 1.    -   2. A secondary substrate consisting of a low-density cotton        fiber is loaded into the bottom void-space 6 of the vessel 1        below the permeable partition 5.    -   3. Example 1 steps 2-4.    -   4. Example 1 step 5, wherein the developing EPM extends through        and around the cotton fiber substrate creating a composite        EPM-cotton fiber material with a higher tensile strength than        the mycelium EPM alone or cotton fiber individually.    -   5. EPM is expanded to a target thickness depending on the depth        of the void space 6 so as to envelope the cotton fiber        adequately, then is separated from the permeable bottom for        post-processing as dictated by the specific application.

The invention thus provides a method of producing a mycologicalmaterial, i.e., a secondary extra-particle fungal matrix, in a simpleinexpensive manner. Further, the invention provides a paradigm toefficiently use a Type II actively aerated static packed-bed bioreactorto manufacture a secondary extra-particle fungal matrix for applicationas a mycological material.

The invention provides a paradigm for production of secondaryextra-particle hyphal matrices (EPM) as:

-   -   A mycological material for replacement of petroleum-based        low-density foams, such as polyurethane foams. The simplified        paradigm described here, as compared to the prior art, provides        an opportunity for direct modification of density and        morphological characteristics of the EPM, increases the        potential scalability of manufacture and material range of        fungal EPM, increasing competitiveness with petroleum-based        foams.    -   A cellular scaffolding, for example the growth of mammalian        cells within the EPM. The described invention is a paradigm for        allowing for specific modification of EPM density and        morphological characteristics by modification of the direct        temperature and gas exchange inputs. This may be applied to        producing EPM specifically intended for providing a scaffold for        mammalian cells for applications such as whole-cut meat        substitutes and biomedical applications. For example, EPM        porosity and density may be specifically modified for        impregnation of mammalian cells of a given size, or the degree        of hyphal agglomeration into cords and directional coherency of        the hyphal cords may be modified to mimic vessels or        vasculature.

1.-11. (canceled)
 12. A mycological material comprising: an aeratedinter-particle hyphal matrix, the aerated inter-particle hyphal matrixconsisting essentially of: a filamentous fungus and a substrate ofdiscrete elements; and an aerated extra-particle hyphal matrix growingfrom the aerated inter-particle hyphal matrix, wherein the aeratedextra-particle hyphal matrix has a higher cell volume density than theaerated inter-particle hyphal matrix, a higher anisotropy than theaerated inter-particle hyphal matrix, and a higher hyphal strandthickness than the aerated inter-particle hyphal matrix.
 13. Themycological material of claim 12, wherein the aerated inter-particlehyphal matrix and the aerated extra-particle hyphal matrix are aeratedwith a preconditioned air.
 14. The mycological material of claim 13,wherein the aerated extra-particle hyphal matrix is positivelygravitropic.
 15. The mycological material of claim 13, wherein theaerated extra-particle hyphal matrix is negatively gravitropic.
 16. Themycological material of claim 13, wherein the preconditioned air ishorizontal through the aerated inter-particle hyphal matrix and theaerated extra-particle hyphal matrix, thereby causing the aeratedextra-particle hyphal matrix to be horizontal and parallel to the flowof the preconditioned air.
 17. The mycological material of claim 13,wherein the preconditioned air comprises one or more of: air, aparamorphogen, a volatile compound, or an aromatic compound.
 18. Themycological material of claim 17, wherein the paramorphogen is one of: aterpene or an alkyl pyrone.
 19. The mycological material of claim 13,wherein the preconditioned air is preconditioned to have a targettemperature and a target humidity.
 20. The mycological material of claim13, wherein the preconditioned air flows laminarly from the aeratedinter-particle hyphal matrix to the aerated extra-particle hyphalmatrix.
 21. The mycological material of claim 12, wherein the aeratedextra-particle hyphal matrix is a target thickness.
 22. The mycologicalmaterial of claim 21, wherein the target thickness is up to 12 inches.23. The mycological material of claim 12, wherein the aeratedinter-particle hyphal matrix is disposed within a bioreactor, whereinthe thickness of the aerated extra-particle hyphal matrix is a functionof the temperature of the bioreactor, a volumetric air flow rate of thebioreactor, an incubation time of the mycological material, and aconstituency of a preconditioned air passing through the bioreactor. 24.The mycological material of claim 12, wherein the discrete elements ofthe substrate are lignocellulosic particles.
 25. The mycologicalmaterial of claim 12, wherein the filamentous fungus is Ganodermatsugae.
 26. The mycological material of claim 12, wherein the aeratedextra-particle hyphal matrix comprises a second substrate.
 27. Themycological material of claim 26, wherein the second substrate is cottonfiber.
 28. A mycological foam, meat substitute or biomedical materialcomprising the mycological material of claim
 12. 29. The biomedicalmaterial of claim 28, wherein the mycological material mimics vessels ofvasculature.
 30. A cellular scaffold comprising the aeratedextra-particle matrix of claim 12.